MicroRNA-CRISPR biosensors for cancer diagnostics

Area of Science:

• Biotechnology
• Molecular Diagnostics
• Biomarker Discovery

Background:

• Circulating microRNAs (miRNAs) are valuable biomarkers for cancer and cardiovascular diseases.
• Low abundance, sequence homology, and matrix effects challenge miRNA detection in biofluids.
• Existing analytical methods require improvement in sensitivity and robustness.

Purpose of the Study:

• To review the latest advances in CRISPR-Cas based microRNA (miRNA) biosensors.
• To evaluate the analytical performance and clinical translational potential of these biosensors.
• To compare CRISPR-based workflows with conventional methods like RT-qPCR and digital PCR.

Main Methods:

• Utilized CRISPR-Cas systems (Cas12a, Cas12b, Cas13a, Cas9) for programmable nucleic acid recognition.
• Employed various signal amplification and readout strategies including fluorescence, electrochemical, and colorimetric methods.
• Critically evaluated analytical performance metrics such as detection limits and specificity.

Main Results:

• CRISPR-based miRNA biosensors achieve femtomolar to attomolar detection limits.
• Demonstrated high specificity and mismatch discrimination capabilities.
• Highlighted versatility across different biofluid samples (plasma, serum, saliva, whole blood, extracellular vesicles).

Conclusions:

• CRISPR-Cas technology presents a highly sensitive and robust platform for miRNA detection.
• These biosensors hold significant potential for minimally invasive clinical diagnostics.
• Standardization, quality control, and clinical validation are crucial for widespread adoption in clinical laboratories.